Immunoassay for compounds of the NBOMe family

Abstract

An immunoassay method for detecting and determining ‘NBOMe’ family designer drugs is described. Also described are components for use in implementing the method, namely, antibodies, detection agents, solid state devices and kits as well as immunogens used to raise the antibodies.

We claim: 
     
       1. An antibody which binds to epitope of Structure (Y) 
       
         
           
           
               
               
           
         
       
       in which R 1  is selected from the group consisting on a halogen, hydrogen, hydroxyl, C 1-3  alkyl, C 1-3  thioalkyl, C 1-3  alkoxy, perfluoromethyl and nitro; R 3  and R 4  are H and R 2  is methoxy;
 wherein the antibody shows a cross-reactivity of 75 to 200% to the epitope of the Structure (Y) and a cross-reactivity of less than 5% to 2-(4-Iodo-2,5-dimethoxyphenyl) ethanamine (2C-I), 2-(4-ethyl-2,5-dimethoxyphenyl)ethanamine (2C-E), 2,5-dimethoxy-4-bromoamphetamine (DOB) and 2-(4-bromo-dimethoxyphenyl)ethanamine (2C-B); and 
 wherein the cross-reactivity is relative to 100% cross-reactivity of the antibody to 2-(4-Iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl] ethanamine (25I-NB)Me). 
 
     
     
       2. The antibody of  claim 1 , which binds to an epitope of at least one of 2-(4-Iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl] ethanamine (25I-NBOMe), 2-(2,5 dimethoxy-4-propylphenyl)-N-(2-methoxybenzyl)ethanamine (25P-NBOMe), Mescaline-[(2-methoxvpheny)methy]ethanamine (Mescaline-NBOMe), 2-(4-Bromo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl] ethanamine (25B-NBOMe), 2-(2,5-Dimethoxy-4-methylphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25D-NBOMe), 2-(4-ethyl-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25E-NBOMe), 2-(2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25H-NBOMe), 2-(2,5-dimethoxy-4-nitrophenyl)-N-(2-methoxybenzyl)ethanamine (25N-NBOMe), 2-[2,5-dimethoxy-4-(-methylsulfanyl)phenyl]-N-(2-methoxybenzyl)ethanamine (25T2-NBOMe), 2-[(4-isopropylsulfanyl)-2,5-dimethoxyphenyl]-N-(2-methoxybenzyl)ethanamine (25T4-NBOMe) and 2-[2,5-dimethoxy-4(propylsulfanyl)phenyl]-N-(2-methoxybenzyl)ethanamine (25T7-NBOMe). 
     
     
       3. The antibody of  claim 1  with an antibody sensitivity of IC50 of 0.2 ng/ml or less for 2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl] ethanamine (25I-NBOMe), 2-(2,5 dimethoxy-4-propylphenyl)-N-(2-methoxybenzyl)ethanamine (25P-NBOMe), mescaline-2-methoxyphenyl)methyl]ethanamine (mescaline-NBOMe), 2 (4-bromo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25B-NBOMe), dimethoxy-4-methylphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25D-NBOMe), 2-(4-ethyl-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25E-NBOMe) 2-(2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25H-NBOMe), 2-(2,5-dimethoxy-4-nitrophenyl)-N-(2-methoxybenzyl)ethanamine (25N-NBOMe), 2-[2,5-dimethoxy-4-(methylsulfanyl)phenyl]-N-(2-methoxybenzyl)ethanamine (25T2-NBOMe), 2-[4-isopropylsulfanyl)-2,5-dimethoxyphenyl]-N-(2-methoxybenzyl)ethanamine (25T4-NBOMe) and 2-[2,5-dimethoxy-4(propylsulfanyl)phenyl]-N-(2-methoxybenzyl)ethanamine (25T7-NBOMe). 
     
     
       4. The antibody of  claim 1  further characterized by being derived from an immunogen of structure II 
       
         
           
           
               
               
           
         
       
       wherein X is a cross-linking group and Y is an antigenicity-conferring carrier. 
     
     
       5. The antibody of  claim 4  wherein X is -(A)-(B)n-, where A is O, S or N and B is a C 1-6  substituted or unsubstituted straight chain alkylene or arylene moiety and n is either 0 or 1. 
     
     
       6. The antibody of  claim 1 , which binds to an epitope of at least one of 25I-NBOMe, 25P-NBOMe, Mescaline-NBOMe, 25B-NBOMe, 25D-NBOMe, 25E-NBOMe, 25H-NBOMe, 25N-NBOMe, 25T2-NBOMe, 25T4-NBOMe and 25T7-NBOMe. 
     
     
       7. The antibody of  claim 1  with an IC50 of 0.2 ng/ml or less for 25I-NBOMe, 25P-NBOMe, Mescaline-NBOMe, 25B-NBOMe, 25D-NBOMe, 25E-NBOMe 25H-NBOMe, 25N-NBOMe, 25T2-NBOMe, 25T4-NBOMe and 25T7-NBOMe. 
     
     
       8. The antibody of  claim 1  further characterized by being derived from an immunogen of structure II 
       
         
           
           
               
               
           
         
       
       wherein X is a cross-linking group and Y is an antigenicity-conferring carrier. 
     
     
       9. A substrate comprising an antibody of one of  claim 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7  or  8 , wherein optionally the substrate is either a microtitre plate or a biochip. 
     
     
       10. A kit comprising an antibody according to one of  claim 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7  or  8 , and optionally one or more detecting agents of structure III 
       
         
           
           
               
               
           
         
       
       in which X is a cross-linking group and Y is a labelling agent which is detectable; and optionally wherein X is -(A)-(B) n1 —, where A is O, S or N and B is a C 1-6  substituted or unsubstituted straight chain alkylene or arylene moiety and  n1  is either 0 or 1; wherein, optionally, X is -(A)-(B) n1 —, where A is O, S or N and B is a C 1-6  substituted or unsubstituted straight chain alkylene terminating, before conjugation with the labelling agent, with a terminal reactive group and  n1  is either 0 or 1; wherein, further optionally, X is -(A)-(B) n1 —, where A is N and B is a C 1-6  substituted or unsubstituted straight chain alkylene terminating, before conjugation with the labelling agent, with a terminal reactive group and  n1  is either 0 or 1.